Evaluation of Therapeutic Potential of Selected Plant-Derived Homeopathic Medicines for their Action against Cervical Cancer

Abstract

Background Plant-derived homeopathic medicines (HMs) are cheap and commercially available but are mechanistically less explored entities than conventional medicines.

Purpose The aim of our study was to evaluate the impact of selected plant-derived HMs derived from Berberis aquifolium (BA), Berberis vulgaris (BV), Mentha piperita (MP), Curcuma longa (CL), Cinchona officinalis (CO), Thuja occidentalis (TO) and Hydrastis canadensis (HC) on cervical cancer (CaCx) cells in vitro.

Methods We screened the mother tincture (MT) and 30C potencies of the above-mentioned HMs for anti-proliferative and cytotoxic activity on human papillomavirus (HPV)-negative (C33a) and HPV-positive CaCx cells (SiHa and HeLa) by MTT assay. Total phenolic content (TPC) and the free-radical scavenging activity of each HM was also determined using standard assays. Phytochemicals reportedly available in these HMs were examined for their potential inhibitory action on HPV16 E6 by in silico molecular docking.

Results All tested MTs induced a differential dose-dependent cytotoxic response that varied with cell line. For C33a cells, the order of response was TO > CL > BA > BV > HC > MP > CO, whereas for SiHa and HeLa cells the order was HC > MP > TO > CO > BA > BV > CL and CL > BA > CO, respectively. 30C potencies of all HMs showed an inconsistent response. Further, anti-CaCx responses displayed by MTs did not follow the order of an HM's phenolic content or free radical scavenging activity. Analysis revealed anti-oxidant content of BA, BV and HC had the lowest contribution to their anti-CaCx activity. Using in silico modeling of molecular docking between the HPV16 E6 protein crystallographic structures (6SJA and 4XR8) and main phytochemical components of BV, BA, HC, CL and TO, their potential to inhibit the HPV16 E6 protein carcinogenic interactions was identified.

Conclusion The study has shown a comparative evaluation of the potential of several plant-derived MTs and HMs to affect CaCx cell line survival in vitro (through cytotoxicity and free radical scavenging) and their theoretical molecular targets in silico for the first time. Data demonstrated that MTs of BA and BV are likely to be the most potent HMs that strongly inhibited CaCx growth and have a strong anti-HPV phytochemical constitution.

Keywords

cervical cancer - human papillomavirus - HPV16 E6 - free-radical scavenging activity - molecular docking

Authors' Contribution

T.S. contributed toward conceptualization, data curation, investigation, methodology, software, validation and roles/writing – original draft. N.A., K.T., A.C., J.Y., T.T., M.J., A.B. and A.K. did the formal analysis and helped in writing original draft. R.H.N., P.G. and A.K. did formal analysis of the resources, writing, review and editing. A.C.B. did the conceptualization, data curation, formal analysis, funding acquisition, investigation, project administration – resources, software, supervision, validation, roles/writing original draft and writing, reviewing, and editing.

All authors read and approved the final manuscript.

Availability of Data and Materials

All data generated or analyzed during this study are included in this published article (and its supplementary information files).

^* Current address: Translational Oncology Laboratory, Department of Zoology, Hansraj College, University of Delhi, Delhi-110007, India.

^** Current address: Department of Zoology, Deshbandhu College, University of Delhi, New Delhi-110019, India.

Publication History

Received: 24 March 2022

Accepted: 12 July 2022

Article published online:
01 March 2023

• References

• 1 Sung H, Ferlay J, Siegel RL. et al. Global Cancer Statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2021; 71: 209-249
• 2 Walboomers JM, Jacobs MV, Manos MM. et al. Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol 1999; 189: 12-19
• 3 Vaccarella S, Laversanne M, Ferlay J, Bray F. Cervical cancer in Africa, Latin America and the Caribbean and Asia: regional inequalities and changing trends. Int J Cancer 2017; 141: 1997-2001
• 4 Yeo-Teh NSL, Ito Y, Jha S. High-risk human papillomaviral oncogenes E6 and E7 target key cellular pathways to achieve oncogenesis. Int J Mol Sci 2018; 19: 19
• 5 Chan WK, Chong T, Bernard HU, Klock G. Transcription of the transforming genes of the oncogenic human papillomavirus-16 is stimulated by tumor promotors through AP1 binding sites. Nucleic Acids Res 1990; 18: 763-769
• 6 Rösl F, Das BC, Lengert M, Geletneky K, zur Hausen H. Antioxidant-induced changes of the AP-1 transcription complex are paralleled by a selective suppression of human papillomavirus transcription. J Virol 1997; 71: 362-370
• 7 Bharti AC, Singh T, Bhat A, Pande D, Jadli M. Therapeutic strategies for human papillomavirus infection and associated cancers. Front Biosci (Elite Ed) 2018; 10: 15-73
• 8 Singh T, Chhokar A, Thakur K. et al. Targeting aberrant expression of STAT3 and AP-1 oncogenic transcription factors and HPV oncoproteins in cervical cancer by Berberis aquifolium . Front Pharmacol 2021; 12: 757414
• 9 Mahata S, Bharti AC, Shukla S, Tyagi A, Husain SA, Das BC. Berberine modulates AP-1 activity to suppress HPV transcription and downstream signaling to induce growth arrest and apoptosis in cervical cancer cells. Mol Cancer 2011; 10: 39
• 10 Mahata S, Maru S, Shukla S. et al. Anticancer property of Bryophyllum pinnata (Lam.) Oken. leaf on human cervical cancer cells. BMC Complement Altern Med 2012; 12: 15
• 11 Mahata S, Pandey A, Shukla S. et al. Anticancer activity of Phyllanthus emblica Linn. (Indian gooseberry): inhibition of transcription factor AP-1 and HPV gene expression in cervical cancer cells. Nutr Cancer 2013; 65 (Suppl. 01) 88-97
• 12 Shukla S, Bharti AC, Hussain S. et al. Elimination of high-risk human papillomavirus type HPV16 infection by ‘Praneem’ polyherbal tablet in women with early cervical intraepithelial lesions. J Cancer Res Clin Oncol 2009; 135: 1701-1709
• 13 Basu P, Dutta S, Begum R. et al. Clearance of cervical human papillomavirus infection by topical application of curcumin and curcumin containing polyherbal cream: a phase II randomized controlled study. Asian Pac J Cancer Prev 2013; 14: 5753-5759
• 14 Protein Data Bank. 6SJA: Structure of HPV16 E6 oncoprotein in complex with IRF3 LxxLL motif. www.rcsb.org/structure/6SJA DOI: 10.2210/pdb6sja/pdb
• 15 Martinez-Zapien D, Ruiz FX, Poirson J. et al. Structure of the E6/E6AP/p53 complex required for HPV-mediated degradation of p53. Nature 2016; 529: 541-545
• 16 USDA. Dr. Duke's Phytochemical and Ethnobotanical Databases. US Department of Agriculture, Agricultural Research Service; 1992-2016
• 17 Samadder A, Das S, Das J, Paul A, Boujedaini N, Khuda-Bukhsh AR. The potentized homeopathic drug, Lycopodium clavatum (5C and 15C) has anti-cancer effect on Hela cells in vitro. J Acupunct Meridian Stud 2013; 6: 180-187
• 18 Bishayee K, Sikdar S, Khuda-Bukhsh AR. Evidence of an epigenetic modification in cell-cycle arrest caused by the use of ultra-highly-diluted gonolobus condurango extract. J Pharmacopuncture 2013; 16: 7-13
• 19 Es S, Kuttan G, Kc P, Kuttan R. Effect of homeopathic medicines on transplanted tumors in mice. Asian Pac J Cancer Prev 2007; 8: 390-394
• 20 Frenkel M, Mishra BM, Sen S. et al. Cytotoxic effects of ultra-diluted remedies on breast cancer cells. Int J Oncol 2010; 36: 395-403
• 21 Karmakar SR, Biswas SJ, Khuda-Bukhsh AR. Anti-carcinogenic potentials of a plant extract (Hydrastis canadensis): I. Evidence from in vivo studies in mice (Mus musculus). Asian Pac J Cancer Prev 2010; 11: 545-551
• 22 Remya V, Kuttan G. Homeopathic remedies with antineoplastic properties have immunomodulatory effects in experimental animals. Homeopathy 2015; 104: 211-219
• 23 Mondal J, Panigrahi AK, Khuda-Bukhsh AR. Anticancer potential of Conium maculatum extract against cancer cells in vitro: Drug-DNA interaction and its ability to induce apoptosis through ROS generation. Pharmacogn Mag 2014; 10 (Suppl. 03) S524-S533
• 24 Abd El-Wahab AE, Ghareeb DA, Sarhan EE, Abu-Serie MM, El Demellawy MA. In vitro biological assessment of Berberis vulgaris and its active constituent, berberine: antioxidants, anti-acetylcholinesterase, anti-diabetic and anticancer effects. BMC Complement Altern Med 2013; 13: 218
• 25 Racková L, Májeková M, Kost'álová D, Stefek M. Antiradical and antioxidant activities of alkaloids isolated from Mahonia aquifolium. Structural aspects. Bioorg Med Chem 2004; 12: 4709-4715
• 26 Müller K, Ziereis K. The antipsoriatic Mahonia aquifolium and its active constituents; I. Pro- and antioxidant properties and inhibition of 5-lipoxygenase. Planta Med 1994; 60: 421-424
• 27 Petruczynik A, Plech T, Tuzimski T. et al. Determination of selected isoquinoline alkaloids from Mahonia Aquifolia; Meconopsis Cambrica; Corydalis Lutea; Dicentra Spectabilis; Fumaria Officinalis; Macleaya Cordata Extracts by HPLC-DAD and Comparison of their Cytotoxic Activity. Toxins (Basel) 2019; 11: 11
• 28 Ungurean C, Carpa R, Câmpean R. et al. Phytochemical and microbial analyses of Berberis sp. Extracts. Rom Biotech Lett 2020; 25: 2132-2139
• 29 Kolluru S, Momoh R, Lin L, Mallareddy JR, Krstenansky JL. Identification of potential binding pocket on viral oncoprotein HPV16 E6: a promising anti-cancer target for small molecule drug discovery. BMC Mol Cell Biol 2019; 20: 30
• 30 Kumar S, Jena L, Sahoo M, Kakde M, Daf S, Varma AK. In silico docking to explicate interface between plant-originated inhibitors and E6 oncogenic protein of highly threatening human papillomavirus 18. Genomics Inform 2015; 13: 60-67
• 31 Zanier K, Charbonnier S, Sidi AO. et al. Structural basis for hijacking of cellular LxxLL motifs by papillomavirus E6 oncoproteins. Science 2013; 339: 694-698